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目的构建旋毛虫感染诱导的小鼠乳腺癌MCF-7细胞基因表达变化的cDNA消减文库,探讨旋毛虫感染诱导的MCF-7细胞基因表达的变化。方法复制MCF-7细胞小鼠模型,设对照组和旋毛虫感染组(实验组),采用抑制性消减杂交技术构建旋毛虫感染诱导的MCF-7细胞上调基因表达消减文库,并利用反向Northernblot技术对所获得基因的特异性表达进行验证。结果成功构建了旋毛虫感染诱导的MCF-7细胞上调基因表达消减文库,克隆的目的基因片段大小分布在200～500bp之间,20个阳性克隆的测序结果经BLAST比对,获得差异表达的已知功能基因8个和未知功能基因4个,这些基因在小鼠乳腺癌细胞中均有表达,而在肌肉组织中无表达。结论旋毛虫感染诱导的MCF-7细胞差异表达基因上调,其中RB基因可能与乳腺癌细胞的生长抑制密切相关,为在基因水平研究旋毛虫抗肿瘤机理奠定了基础。
Objective To construct cDNA subtractive library of mouse breast cancer MCF-7 cells induced by Trichinella spiralis infection and to investigate the gene expression changes of MCF-7 cells induced by Trichinella spiralis infection. Methods MCF-7 cells were induced by Trichinella spiralis infection in MCF-7 cells. The control group and Trichinella spiralis infection group (experimental group) were established. Suppression subtractive hybridization (SSH) The technique validates the specific expression of the obtained genes. Results The cDNA library of MCF-7 cells induced by Trichinella spiralis infection was successfully constructed. The cloned target gene fragments were distributed between 200 and 500bp in size. The positive results of 20 positive clones were BLAST-aligned and the differentially expressed Eight genes were known and four were unknown genes. These genes were expressed in mouse breast cancer cells but not in muscle tissue. Conclusion Trichinella spiralis infection-induced MCF-7 cells differentially expressed genes were up-regulated, of which RB gene may be closely related to the growth inhibition of breast cancer cells in order to study the anti-tumor mechanism of Trichinella at the genetic level.